Kwak Dong Hoon, Seo You Na, Lee Ju Hyoung, Park Soon Ju, Cho Young Ho, Kim Ji-Su, Kim Sun-Uk, Choo Young-Kug
Institute of Glycosciences, Wonkwang University, Iksan, Republic of Korea.
Institute of Aribio, Sungnam, Republic of Korea.
Anim Cells Syst (Seoul). 2018 Mar 22;22(3):157-164. doi: 10.1080/19768354.2018.1453546. eCollection 2018.
Pig-human xenotransplantation can trigger cell-mediated immune responses. We explored the role of gangliosides in inflammation related to immune rejection in xenotransplantation. Co-culture of xenogeneic cells (pig-MSCs and RAW264.7) was used to emulate xenotransplantation conditions. MTT assay results indicated that cell viability was significantly decreased in pADMSCs co-cultured with RAW264.7 cells. GM1 and GM3 were highly expressed in pADMSCs co-cultured with RAW264.7 cells. pADMSCs co-cultured with RAW264.7 cells strongly expressed pro-inflammatory proteins such as COX-2, iNOS, p50, p65, pIκBα, and TNF-α. GM1-knockdown pADMSCs co-cultured with RAW 264.7 cells did not show significantly altered cell viability, but pro-inflammatory proteins were markedly inhibited. Co-culture of pADMSCs with RAW264.7 cells induced significant phosphorylation (p) of JNK1/2 and pERK1/2. However, pERK1/2 and pJNK1/2 were decreased and MEK1/2 and Raf1 were suppressed in GM1-knockdown pADMSCs co-cultured with RAW264.7 cells. Thus, the Raf-1/MEK1/2/ERK1/2 and JNK1/2 pathways were significantly upregulated in response to increases of GM1 in co-cultured xenogeneic cells. However, the inflammatory response was suppressed in co-culture of GM1-knockdown pADMSCs with RAW264.7 cells via down-regulation of the Raf-1/MEK1/2/ERK1/2 and JNK1/2 pathways. Therefore, the ganglioside GM1 appears to play a major role in the inflammatory response in xenotransplantation via the Raf-1/MEK1/2/ERK1/2 and JNK1/2 pathways.
猪-人异种移植可引发细胞介导的免疫反应。我们探讨了神经节苷脂在异种移植中与免疫排斥相关的炎症中的作用。采用异种细胞（猪间充质干细胞和RAW264.7）共培养来模拟异种移植条件。MTT分析结果表明，与RAW264.7细胞共培养的猪脂肪来源间充质干细胞（pADMSCs）的细胞活力显著降低。GM1和GM3在与RAW264.7细胞共培养的pADMSCs中高表达。与RAW264.7细胞共培养的pADMSCs强烈表达促炎蛋白，如COX-2、iNOS、p50、p65、pIκBα和TNF-α。与RAW 264.7细胞共培养的GM1敲低pADMSCs的细胞活力没有显著改变，但促炎蛋白受到明显抑制。pADMSCs与RAW264.7细胞共培养诱导JNK1/2和pERK1/2显著磷酸化（p）。然而，在与RAW264.7细胞共培养的GM1敲低pADMSCs中，pERK1/2和pJNK1/2降低，MEK1/2和Raf1受到抑制。因此，在共培养的异种细胞中，Raf-1/MEK1/2/ERK1/2和JNK1/2通路因GM1增加而显著上调。然而，通过下调Raf-1/MEK1/2/ERK1/2和JNK1/2通路，GM1敲低pADMSCs与RAW264.7细胞共培养时炎症反应受到抑制。因此，神经节苷脂GM1似乎通过Raf-1/MEK1/2/ERK1/2和JNK1/2通路在异种移植的炎症反应中起主要作用。
